Quantitative and Wide-Ranging Profiling of Phospholipids in Human Plasma by Two-dimensional Liquid Chromatography/Mass Spectrometry

• Published in: Analytical chemistry (Washington) Vol. 82; no. 23; pp. 9858 - 9864
• Main Authors: Sato, Yoshiaki, Nakamura, Tatsuji, Aoshima, Ken, Oda, Yoshiya
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 01.12.2010
• Subjects: Alzheimer Disease - blood; Alzheimer Disease - metabolism; Alzheimer's disease; Analytical chemistry; Animals; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography; Chromatography, High Pressure Liquid - methods; Discriminant Analysis; Exact sciences and technology; Humans; Least-Squares Analysis; Lipids; Male; Mass spectrometry; Other chromatographic methods; Phosphatidylserines - blood; Phospholipids - blood; Plasma; Rats; Rats, Sprague-Dawley; Software; Spectrometric and optical methods; Spectrometry, Mass, Electrospray Ionization - methods; Solid phase extraction; Plasma; Peak resolution; Two dimensional chromatography; Alzheimer disease; Liver; Phospholipid; Lipids; Standard; Liquid phase; Chemical enrichment; Characterization; Ion trap; Sample preparation; Quantitative analysis; Human; Discriminant analysis; Sample; Use; Data processing; Liquid chromatography; Method; PLS regression; Software; Mass spectrometry
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac102211r

Normal-phase or reverse-phase liquid chromatography has been used in phospholipidomics for lipid separation prior to mass spectrometry analysis. However, separation using a single separation mode is often inadequate, as high-abundance phospholipids can mask large numbers of low-abundance lipids of interest. In order to detect and quantify low-abundance phospholipids, we present a novel two-dimensional (2D) approach for sensitive and quantitative global analysis of phospholipids. The methodology monitors individual glycerolipids and phospholipids through the use of a new quantitative normal-phase, solid-phase extraction procedure, followed by molecular characterization and relative quantification using an ion-trap Orbitrap equipped with a reverse-phase liquid chromatograph, with data processing by MS++ software. The CV (%) of the peak area of each lipid standard was less than 15% with this extraction method. When the method was applied to a liver sample, we could detect more phosphatidylserine (PS) compared to the previous method. Finally, our developed method was applied to Alzheimer’s disease (AD) plasma samples. Several hundred peaks were detected from a 60 μL plasma sample. A partial-least-squares discriminant analysis (PLS-DA) plot using peak area ratio gave a unique group of PLS scores which could distinguish plasma samples of Alzheimer’s disease (AD) patients from those of age-matched healthy controls.